Development and Evaluation of Quality Control Dried Blood Spot Materials in Newborn Screening for Lysosomal Storage Disorders
Development and Evaluation of Quality Control Dried Blood Spot Materials in Newborn Screening for Lysosomal Storage Disorders
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DOI：
10.1373/clinchem.2008.111864
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发表时间：
2009-01-01
影响因子：
9.3
通讯作者：
Hannon, W. Harry
中科院分区：
文献类型：
--
作者：
De Jesus, Victor R.;Zhang, X. Kate;Hannon, W. Harry
背景技术背景：溶酶体胆积症（LSD）包括40多种遗传疾病，其导致通常会被溶酶体酶降解的产物的积累。一种基于串联质谱（MS/MS）的方法可用于新生儿筛查5个LSD，许多实验室正在启动试点研究，以评估将该方法纳入其筛查小组。我们开发和评估干血斑（DBS）的LSDs的QC材料，并使用MS/MS方法，以探讨其适用性LSD QC monitoring.METHODS：我们培养3.2毫米从DBS控制冲孔20-24小时与含有底物和内标的检测鸡尾酒。使用MS/MS，我们对所得产物和内标物进行定量。样品一式三份连续运行3天，结果报告为产品与内标物的比值和酶活性单位（μ mol/L/h）。高、中和低系列的酶活性日间不精密度（CV）分别为：半乳糖苷α-半乳糖苷酶3.4%-14.3%，酸性α-半乳糖苷酶A 6.8%-24.6%，酸性鞘磷脂酶为7.36%~ 22.1%，酸性α-葡萄糖苷酶为6.2%~ 26.2%，溶酶体酸性α-葡萄糖苷酶为7.0%~ 24.8%（n = 9）。此外，在-20 ℃和4 ℃下储存的DBS在187天的时间内表现出最小的酶活性损失。DBS存储在37度和45摄氏度有较低的活性值超过187天的evaluationtime.CONCLUSIONS：合适的QC材料新生儿筛选LSDs进行DBS LSD筛选的实验室开发。观察到良好的材料线性，拟合优度值为0.953及更高。QC材料可由通过MS和/或更常规的基于荧光的筛选方法进行LSD分析的筛选实验室使用。(C)2008年美国临床化学协会
BACKGROUND: Lysosomal storage disorders (LSDs) comprise more than 40 genetic diseases that result in the accumulation of products that would normally be degraded by lysosomal enzymes. A tandem mass spectrometry (MS/MS)-based method is available for newborn screening for 5 LSDs, and many laboratories are initiating pilot studies to evaluate the incorporation of this method into their screening panels. We developed and evaluated dried blood spot (DBS) QC materials for LSDs and used the MS/MS method to investigate their suitability for LSD QC monitoring.METHODS: We incubated 3.2-mm punches from DBS controls for 20-24 h with assay cocktails containing substrate and internal standard. Using MS/MS, we quantified the resulting product and internal standard. Samples were run in triplicate for 3 consecutive days, and results were reported as product-to-internal standard ratios and enzyme activity units (mu mol/L/h).RESULTS: Enzyme activity interday imprecision (CV) for the high, medium, and low series were 3.4%-14.3% for galactocerebroside a-galactosidase, 6.8%-24.6% for acid a-galactosidase A, 7.36%-22.1% for acid sphingomyelinase, 6.2%-26.2% for acid alpha-glucocere-brosidase, and 7.0%-24.8% for lysosomal acid a-glucosidase (n = 9). In addition, DBS stored at -20 degrees and 4 degrees C showed minimal enzyme activity loss over a 187-d period. DBS stored at 37 degrees and 45 degrees C had lower activity values over the 187-day evaluation time.CONCLUSIONS: Suitable QC materials for newborn screening of LSDs were developed for laboratories performing DBS LSD screening. Good material linearity was observed, with goodness-of-fit values of 0.953 and higher. The QC materials may be used by screening laboratories that perform LSD analysis by MS and/or more conventional fluorescence-based screening methods. (C) 2008 American Association for Clinical Chemistry